F lusiTex
Fluorescence sensing integrated into medical textiles
Current Wound Monitoring
Wound healing involves a complex series of biochemical events. Though our
understanding of the healing process has improved, we still use low tech dressings.
Current Methods
Full or partial removal of wound pad
Visual Observation
Skin Irritation
Increased chances of infection
Only qualitative information
Collecting biochemical information
Highly invasive
Expensive
The Business Case
$12bn worldwide market aimed solely at wound care
Wound monitoring consists less than 1% of this
industry
6 weeks treatment of a chronical wound
35,000 CHF
Benefits in sensing the wound include
Reduce hospitalization time
Provide better treatment
Prevents amputation
Working Principle
We want to build a wound healing system that can monitor the healing
process and providing quantitative data.
Non-invasive
TOF camera
One fluorescence signal
per sensing parameter
per wound area
Sensing layer
pH,
metabolites,
oxygen,
Wound
Pad Structure
material
proteins and enzymes
E
E
metabolites and enzymes,
H + , O2
Several detection spots on the pad (microfabricated):
pH
Oxygen
Various metabolites / enzymes
etc.
The Team
Dr. Luciano Boesel
Sensing chemistry and matrix development
Dr. Greta Faccio
Enzyme engineering and coupling
Dr. Stefano Cattaneo
Fluorescent lifetime camera development
Prof. Bradley Nelson
Coating/microfabrication of wound pad
Prof. Brigitte von Rechenberg
In vivo evaluation of wound pad
Wound parameters
pH Glucose
Depends on time course
Bio-marker to assess the
and wound-stage.
Normal healing -> pH Oxygen wound status.
Infected wounds:
increases during
granulation and concentration of around
decreases afterwards to 0.31.0 mM.
reach a value of 4-6. 1) Controls cell Non-infected wounds:
Impaired healing -> proliferation concentration of around
2) Anti-bacterial 5.07.6 mM.
the pH oscillates between
activities
pH 7-8
3) Creating new blood
vessels
(angiogenesis),
which is essential to
the growth and
survival of repaired
The sensors
Non-invasive
TOF camera
One fluorescence signal
per sensing parameter
per wound area
Sensing layer
Parameter
Wound
Fluorescent pH-marker
5(6)-Carboxynaphthofluorescein Fluorescence intensity / a.u.
700
pH = 8
600 pH =
7.7
500 pH =
7.3
400 pH = 7
pH =
300 6.7
200 pH =
6.3
100 pH = 6
pH =
0 5.5
620 720 pH = 5
Wavelength / nm
700
Fluorescence Intensity / a.u.
600
500
400
300
200
100
0
5 5.5 6 6.5 7 7.5 8
pH
Enzimatic glucose sensing
Principle of glucose sensor functioning Storage and stability of funtionalized matrix
Glucose oxidase HRP, 30C
Glucose + O2 Gluconolactone + H2O2
Horseradish peroxidase
H2DCF + H2O2 DCF + H2O
Sensors after immobilization on alginate
Reaction with glucose solutions
GOx, 30C
glucose 0 0.02 0.5 2 5 10 [mM]
Non-enzymatic glucose sensing
Oxygen Sensors Fabrication
Ethyl cellulose
PtOEP
1 cm
Alginate gel
Long Luminescence decay times
Luminescence brightness of the probe
Can be detected @ low
concentration
Solubility in the polymeric matrices
Chemical stability and photostability
Commercial or synthetic availability
Oxygen Sensor System
In the presence of oxygen the lifetime of PtOEP is quenched
Advantage of fluorescence lifetime vs intensity:
independent of dye concentration, excitation light fluctuations, etc.
Controlling sensors positions
Currently we are also working to precisely position the sensors in the gel
matrix.
Antinode
Top view PZT Reflector
Pressure
Node node
/2
UV excitation
Sensors
Side view
PZT 15
m
The Lifetime Camera
Non-invasive
TOF camera
One fluorescence signal
per sensing parameter
per wound area
Sensing layer
pH,
metabolites,
oxygen,
Wound
Fluorescence lifetime imaging
Fluorescence lifetime: the time at which the
intensity has decayed to 1/e of the original
value.
FlusiTex FLIM prototype:
Compact, cost-effective,
simple
Standalone system with
integrated light source
Suitable for decays from nano-
to microseconds
Conventional FLIM
setup:
Bulky
Complex
Expensive
MATLAB GUI Software
Phase-resolved
intensity images
Phase shift Fluorescence
Lifetime (ns)
FLIM Oxygen Measurements
Ocean Optics RedEye
0 50 % Oxygen Sensing Patch 100
% %
3000
2500
Lifetime [ns]
2000
1500
1000
500
0 10 20 30 40 50 60 70 80 90 100
Oxygen O2 Concentration [%]
Testing in Animal Models
Lactate
pH
Glucose
O2
Pig Pig
Aim 1: testing fresh wounds Aim 2: testing infected wounds
Feasibility for wound sensor Feasibility of primary closure
pad Monitoring over time
Feasibility of parameters
Wound model Wound model
Aim 1: Standardized wound on Aim 2: Standardized wound on
dorsum of animal; size 4 x 4cm2; tibia directly above bone; size 4 x
comparison wound pad to 4cm2; infection with S. aureus;
standard wound bandage, comparison wound pad to
histology of wound healing standard wound bandage
https://www.eqalix.com/technology-and-pip
WP5 : In Vivo Evaluation
Standardized deep wounds
Dorsum of mini-pigs
4 deep wounds, ~16cm2 each (<1%TBSA)
Standardized depth with dermatome
Treatment groups:
Group 1: Standard of Care
Group 2: Flusitex without molecular sensors (scaffold only)
4 cm
Group 3: Flusitex dressing
4 cm
Wound area
Imaging area
6mm diameter
WP5 : In Vivo Evaluation
Treatment and tracking timeline
Endpoint: Wound closure or Day 21
Few dressing changes. Daily imaging.
Imaging border zones and center of each wound dressing
Wound imaging parameters (pO2, pH, metabolites)
Wound healing (Epithelialisation, wound surface area, granulation tissue)
Proof of concept: Infected wounds
Inoculation with Pseudomonas Aeruginosa at Day 2
Tracking of infection parameters (local swabs, systemic markers)
Antibiotic therapy after infection establishment
Same treatment distribution as non-infected animals
Achievements and Future Efforts
Achievements
Development of:
o pH sensor
o biomarker sensors
o oxygen sensor
Coupling or integration of the sensor systems to/into the coating matrix
Development of hand-held prototype for fluorescence lifetime imaging in
the nanosecond range
Future Efforts
Microfabrication of the functional matrix on a wound pad
In vitro and in vivo analysis of the monitor pads
Outcome - Gateways
FlusiGate
Flawa, Kenzen, Schoeller,
TheranOptics:
Project to develop a ratiometric
sensor for wound monitoring
based on pH responsive dyes
and a point fluorescent detector
FlusiSafe
U-NICA Solutions AG : project to
investigate the potential of the
FLIM imager for anti-
counterfeiting & product
protection
Prof. Bradley Nelson
Dr. Luciano Boesel
Dr. Daniel Ahmed
Dr. Claudio Toncelli
Dr. Chen Xiangzhong
Dr. Alina Osypova
Dr. Guido Panzarasa
Thanks!
Dr. Stefano Cattaneo
Dr. Greta Faccio Mr. Christoph Hofer
Dr. Dagmara Jankowska Dr. Hans-Rudolf Graf
Dr. Bernhard Schaffer
Prof. Brigitte von Rechenberg
Dr. Salim Darwiche
